Polymerase chain reaction (PCR) for detection of pathogenic microorganisms in bacteriological monitoring of dairy products

1995 ◽  
Vol 146 (1) ◽  
pp. 85-97 ◽  
Author(s):  
M Allmann ◽  
C Höfelein ◽  
E Köppel ◽  
J Lüthy ◽  
R Meyer ◽  
...  
2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.


2012 ◽  
Vol 66 (9) ◽  
Author(s):  
Kristýna Turková ◽  
Bohuslav Rittich ◽  
Alena Španová

AbstractSeveral DNA amplification-based methods were used for identification and evaluation of the relation between lactobacilli isolated from breastfed full-term infant faeces (31 strains), dairy products (5 strains) and silage (1 strain). Twenty-seven strains isolated from infant faeces were identified as Lactobacillus rhamnosus (9), Lactobacillus gasseri (6), Lactobacillus paracasei (4), Lactobacillus fermentum (4), Lactobacillus salivarius (2), Lactobacillus plantarum (1), and Lactobacillus helveticus (1) using 10 species-specific polymerase chain reactions (PCRs), multiplex PCR for the Lactobacillus casei group, and sequencing of 16S rDNA. Four strains were not identified. Six strains isolated from dairy products and silage were identified as Lactobacillus rhamnosus. A repetitive extragenic palindromic polymerase chain reaction (rep-PCR) with primer (GTG)5 and a randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) with primer M13 were used for confirmation of species identification. Fingerprints were used for evaluation of the relatedness of lactobacilli. Differences between strains from infant faeces, dairy products, and silage were not detected.


1996 ◽  
Vol 59 (10) ◽  
pp. 1031-1036 ◽  
Author(s):  
MARYANNE DRAKE ◽  
CHRISTOPHER L. SMALL ◽  
KEMET D. SPENCE ◽  
BARRY G. SWANSON

Species-specific primers for use in the polymerase chain reaction (PCR) were designed to differentially amplify DNA from the common dairy lactobacillus species Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus helveticus, and Lactobacillus acidophilus. A method for rapid extraction of bacterial DNA from dairy products was developed. The sensitivity of bacterial DNA extraction from food and subsequent amplification by PCR was 100 cells total. Lactobacillus DNA was extracted and identified from commercial yoghurts, acidophilus milk, and cheeses. The methodology allows the presumptive identification of dairy lactobacilli in less than 6 hours.


2021 ◽  
pp. 12-14
Author(s):  
Николай Анатольевич Жижин

Идентификация пищевого сырья, применяемого для производства продуктов питания, является важным аспектом безопасности здоровья человека. Развитие аллергических реакций, непереносимость отдельных компонентов пищи и т. д. являются поводом для вынесения информации о составе пищевой продукции на этикеточную надпись. Также к важным факторам оценки продуктов питания можно отнести и видовую «чистоту». Этот показатель необходимо учитывать не только в качестве выявления фальсификации продукции более дешевым сырьем, но и для безопасности конечного потребителя. Для идентификации видового состава молока и молочной продукции достаточно успешно используется метод полимеразной цепной реакции. Развитие этого метода привело к появлению различных подходов его использования. Одним из них является метод мультиплексной полимеразной реакции, который позволяет одновременно проводить процесс амплификации различных последовательностей ДНК. Что позволяет использовать данный метод для одновременного определения двух и более видов сельскохозяйственных животных в течение проведения одного ПЦР-анализа. В данной работе показаны возможности применения мультиплексной ПЦР для идентификации молока и молочной продукции. Показано, что в течение одного анализа определяются специфические праймеры, характерные для трех видов животных: коровы, овцы и козы. Применяемая методика также была использована на молочной продукции, прошедшей термическую обработку, в результате чего установлена возможность использования мультиплексной ПЦР для анализа такой продукции. Предел обнаружения при проведении ПЦР-анализа составил 0,1 %. The identification of food raw materials used for food production is an important aspect of human health safety. Development of allergic reactions, intolerance to certain food components, etc. are the reason for placing information on the composition of food products on the label inscription. Species «purity» can also be attributed to the important factors in assessing food products. This indicator must be taken into account not only as a detection of product counterfeiting with cheaper raw materials, but also for the safety of the end consumer. To identify the species composition of milk and dairy products, the method of polymerase chain reaction is quite successfully used. The development of this method has led to the emergence of various approaches to its use. One of which is the multiplex polymerase reaction method, which allows simultaneous amplification of various DNA sequences. That allows you to use this method for the simultaneous determination of two or more species of farm animals during one PCR analysis. In this work, the possibilities of using multiplex PCR for the identification of milk and dairy products were shown. It was shown that during one analysis, specific primers characteristic of three species of animals: cow, sheep and goat are determined. The applied technique was also used on heat-treated dairy products, as a result of which the possibility of using multiplex PCR for the analysis of such products was established. The detection limit for PCR analysis was 0.1 %.


2020 ◽  
Vol 73 (2) ◽  
pp. 380-383
Author(s):  
Pavlo V. Fedorych ◽  
Gennadiy I. Mavrov ◽  
Tetiana V. Osinska ◽  
Yuliia V. Shcherbakova

The aim was to perform systematic review of genitourinary protozoan invasion and analyze their pathogenicity and the ability to influence the genitourinary infections. Materials and methods: For systematic review of papers the EMBASE and PubMed databases were searched. We also reviewed our own pilot studies using real-time polymerase chain reaction (PCR) to determine Trichomonas tenax, Pentatrichomonas hominis and Giardia lamblia. Conclusions: Trichomonas tenax, Pentatrichomonas hominis, Giardia lamblia can cause genitourinary invasion in addition to Trichomonas vaginalis. Their eradication is obligatory at least for not keeping intact pathogenic microorganisms phagocyted by Trichomonas spp. Defining the protozoan forms is important in preventing of genital infections recurrences and reinfections.


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